Unknown,Transcriptomics,Genomics,Proteomics

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RNA Seq analysis of Mefs Isolated from STC1+/+ and STC1-/- mice


ABSTRACT: Evaluation of X-linked gene expression in MEFs isolated from STC1 wild type and knock out mice X chromosome inactivation (XCI) is initiated in cis by the Xist RNA, which coats the inactive X chromosome (Xi) from which it is produced. We performed a large-scale RNA interference screen to identify trans-acting XCI factors (XCIFs) that comprise regulators of cell signaling and transcription. We find that the XCIFs promote Xist expression and/or localization to the Xi. One of the XCIFs, STC1, is a glycoprotein found in both the cytoplasm and nucleus and whose function is poorly understood. A homozygous mouse knockout of STC1 has a defect in XCI but surprisingly is phenotypically normal. We performed transcriptome profiling (RNA-Seq) experiments to determine whether the expression levels of X-encoded genes were elevated in Stc1-/- female MEFs. In these experiments, RNA was prepared from three independent cultures of Stc1+/+ or Stc1-/- female MEFs. RNA samples were processed and amplified followed by deep sequencing. The similarity of X-linked gene expression between Stc1+/+ and Stc1-/- MEFs was statistically significant. The vast majority of autosomal genes were also expressed at comparable levels in Stc1+/+ and Stc1-/- MEFs. Sequenced mRNA isolated from the STC1+/+ or STC1-/- MEFs.

ORGANISM(S): Mus musculus

SUBMITTER: Sanchita Bhatnagar 

PROVIDER: E-GEOD-47395 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Genetic and pharmacological reactivation of the mammalian inactive X chromosome.

Bhatnagar Sanchita S   Zhu Xiaochun X   Ou Jianhong J   Lin Ling L   Chamberlain Lynn L   Zhu Lihua J LJ   Wajapeyee Narendra N   Green Michael R MR  

Proceedings of the National Academy of Sciences of the United States of America 20140818 35


X-chromosome inactivation (XCI), the random transcriptional silencing of one X chromosome in somatic cells of female mammals, is a mechanism that ensures equal expression of X-linked genes in both sexes. XCI is initiated in cis by the noncoding Xist RNA, which coats the inactive X chromosome (Xi) from which it is produced. However, trans-acting factors that mediate XCI remain largely unknown. Here, we perform a large-scale RNA interference screen to identify trans-acting XCI factors (XCIFs) that  ...[more]

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